GRIBCG

GRIBCG identifies single guide RNAs (sgRNAs) for Streptococcus pyogenes Cas9 (SpCas9) to design balancer chromosomes by selecting multiple chromosomal cut sites while minimizing off-target effects.

Key Features:

• sgRNA identification for SpCas9: Identifies single guide RNAs (sgRNAs) compatible with Streptococcus pyogenes Cas9.
• Multi-site targeting: Selects sgRNAs that enable efficient cutting at multiple sites on a target chromosome to facilitate chromosomal inversions or rearrangements.
• Off-target minimization: Prioritizes sgRNAs to minimize off-target cleavage elsewhere in the genome.
• Cross-species validation: sgRNA selection approach validated across six model organisms.
• Benchmarking: Performance benchmarked against two routinely used Drosophila balancer chromosomes.

Scientific Applications:

• Balancer chromosome generation: Design of chromosomal inversions and rearrangements to suppress meiotic recombination and maintain specific gene combinations.
• Cross-species engineering: Application of CRISPR/Cas9-mediated inversion design to multicellular organisms beyond Drosophila, including biomedical and plant genetics contexts.
• Genetic maintenance and mapping: Enabling maintenance of allelic combinations and facilitation of genetic studies that require stable chromosomal configurations.

Methodology:

Systematic computational selection of sgRNAs for SpCas9 optimizing efficient multi-site chromosomal cutting and off-target minimization, with validation across six model organisms and benchmarking against two Drosophila balancer chromosomes.